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Fig. 3 | Microbial Cell Factories

Fig. 3

From: Use of the KlADH3 promoter for the quantitative production of the murine PDE5A isoforms in the yeast Kluyveromyces lactis

Fig. 3

Analysis of ADH and PDE5 activities from cellular extracts. a In-gel native ADH pattern from the wild type (WT) and pα-PDE5A1 transformed strain. Cells were grown in YPG or YPD medium for 1–5 days. The migrating position of ADH isoforms is reported on the left. b cGMP hydrolysing activities from extracts of WT and pα-PDE5A1 transformed strain. Extracts were obtained from YPG-grown cultures of a. c Western blot of PDE5A1 from the 2–4 days YPD- YPG-grown cultures extracts of a (lanes 7–12) and extracts from YP lactate or acetate cultures. d cGMP hydrolysing activities with/without 200 nM Sildenafil (sild). Soluble (s) and pellet fraction (p) were from the 4 days YPG-grown cultures extracts of a. e Western blot of PDE5A1 from the 4 days YPG-grown cellular extracts and medium supernatants of a concentrated 20 times. N18TG2 (N18) murine neuroblastoma cell line extract is used as PDE5 standard. Each lane contains 10 μg of protein extract for ADH analysis, 30 μg for western blot of PDE or the concentrated equivalent of 0.3 mL of medium supernatant. Arrows indicate the migrating position of PDE5 bands

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